Hodorovich Dana R, Fryer Harris Tiara, Burton Derek, Neese Katie, Bieler Rachael, Chudasama Vimal, Marsden Kurt C
Department of Biological Sciences, North Carolina State University, North Carolina, United States of America.
Current Address: National Institute of Environmental Health Sciences, Durham, North Carolina, United States of America.
bioRxiv. 2023 Aug 1:2023.07.31.551330. doi: 10.1101/2023.07.31.551330.
The larval zebrafish is a highly versatile model across research disciplines, and the expanding use of behavioral analysis has contributed to many advances in neuro-psychiatric, developmental, and toxicological studies, often through large-scale chemical and genetic screens. In the absence of standardized approaches to larval zebrafish behavior analysis, however, it is critical to understand the impact on behavior of experimental variables such as the size of testing arenas and the choice of embryo medium. Using a custom-built, modular high-throughput testing system, we examined the effects of 4 testing arena sizes and 11 types of embryo media on conserved sensorimotor behaviors in zebrafish larvae. Our data show that testing arena size impacts acoustic startle sensitivity and kinematics as well as spontaneous locomotion and thigmotaxis, with fish tested in larger arenas displaying reduced startle sensitivity and increased locomotion. We also find that embryo media can dramatically affect startle sensitivity, kinematics, habituation, and pre-pulse inhibition, as well as spontaneous swimming, turning, and overall activity. Common media components such as methylene blue and high calcium concentration consistently reduced startle sensitivity and locomotion. To further address how the choice of embryo medium can impact phenotype expression in zebrafish models of disease, we reared mutant larvae, a model of CHARGE syndrome with previously characterized morphological and behavioral phenotypes, in 5 different types of media and observed impacts on all phenotypes. By defining the effects of these key extrinsic factors on larval zebrafish behavior, these data can help researchers select the most appropriate conditions for their specific research questions, particularly for genetic and chemical screens.
斑马鱼幼体是一种适用于多个研究领域的通用模型，行为分析的广泛应用常常通过大规模化学和基因筛选，推动了神经精神病学、发育学和毒理学研究的诸多进展。然而，由于缺乏标准化的斑马鱼幼体行为分析方法，了解诸如测试场地大小和胚胎培养基选择等实验变量对行为的影响至关重要。我们使用定制的模块化高通量测试系统，研究了4种测试场地大小和11种胚胎培养基对斑马鱼幼体保守感觉运动行为的影响。我们的数据表明，测试场地大小会影响听觉惊吓敏感性、运动学以及自发运动和趋触性，在较大场地中测试的鱼表现出较低的惊吓敏感性和较高的运动能力。我们还发现，胚胎培养基会显著影响惊吓敏感性、运动学、习惯化和前脉冲抑制，以及自发游泳、转向和整体活动。诸如亚甲蓝和高钙浓度等常见培养基成分会持续降低惊吓敏感性和运动能力。为了进一步探讨胚胎培养基的选择如何影响疾病斑马鱼模型中的表型表达，我们在5种不同类型的培养基中饲养了突变幼体（一种患有CHARGE综合征的模型，具有先前已表征的形态和行为表型），并观察到对所有表型的影响。通过确定这些关键外部因素对斑马鱼幼体行为的影响，这些数据可以帮助研究人员为其特定研究问题选择最合适的条件，特别是在基因和化学筛选方面。
